靶向TRPV1受体的蜈蚣毒素环肽cRhTx的合成及活性研究

Synthesis and biological activity study of cyclized centipede toxin cRhTx against TRPV1

目的合成首尾环化的蜈蚣毒素环肽(cRhTx),评价它对TRPV1配体门控离子通道的影响。方法采用Fmoc-固液相联用多肽合成技术和反相高效液相色谱-质谱联用技术制备野生型蜈蚣毒素(RhTx)与cRhTx,利用圆二色谱法分析两者的结构特点,并进一步采用电生理学法检测两者对TRPV1受体激活的影响。结果首次通过引入GGAAGG柔性连接片段合成了结构新颖的cRhTx,cRhTx和RhTx具有较高的结构相似性,且cRhTx对TRPV1受体的激动作用与RhTx相似。结论引入GGAAGG柔性连接片段,对野生型RhTx进行环化,可以保持其对TRPV1的激动作用,获得的cRhTx具有重要应用潜力。

Objective N,C-termini cyclized centipede toxin RhTx(cRhTx)was synthesized,and the structural difference between the cRhTx and the wild-type RhTx was compared,and the activating difference of the TRPV1 ligand-gated ion channel was analyzed.Methods cRhTx and wild-type RhTx were synthesized by Fmoc-SPPS and LC-MS.Structures of cRhTx and wild-type RhTx were analyzed by circular dichroism.Electrophysiological methods were used to compare the activation of TRPV1 between cRhTx and wild-type RhTx.Results cRhTx was successfully synthesized firstly through grafting linker(GGAAGG).CD spectra showed structural similarity between cRhTx and wild-type RhTx.The cRhTx had a slightly stronger agonistic effect on TRPV1 than RhTx.Conclusion Introduction of GGAAGG flexible ligation fragment,cyclization of wild-type RhTx,could maintain the agonistic effect on TRPV1,and the finally obtained cRhTx has great application potential.